Applying high-resolution single particle cryoEM to a broad range of macromolecular complexes requires both advanced technologies that deliver images with sufficient high quality as well as homogeneous samples with sufficient structural features for image alignment. We developed method based on novel hardware of direct electron counting camera to correct motion-induced image blurring, which has been a limiting factor for routinely achieving high-resolution structure determination by single particle cryoEM. We also developed biochemical method to generate and to screen for monoclonal conformational Fabs against small target proteins to facilitate image alignment of small proteins that do not contains sufficient structural features for image alignment. By combining the approaches of based on both hardware and biochemistry, we are attempting achieving high-resolution single particle cryoEM for a broad range of proteins that are smaller in size and lower in symmetry than icosahedral viruses.

Attempting high-resolution single particle cryoEM of macromolecular complexes: Combining hardware and biochemistry

245th National Meeting (2013)

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